Sulfohydroxamic acids and sulfohydroxamates and their use as MEK inhibitors

ABSTRACT

Sulfohydroxamic acid diarylamines and derivatives thereof; compositions made from them; and methods of using them.

This application is a Divisional application U.S. Ser. No. 09/889,101filed Jul. 11, 2001, now U.S. Pat. No. 6,455,582, which is a 371application of PCT 99/30417 filed Dec. 21, 1999, which claims thebenefit of priority to U.S. provisional application Ser. No. 60/115,652filed Jan. 13, 1999 and U.S. provisional application Ser. No. 60/122,417filed Mar. 2, 1999.

The invention relates to sulfohydroxamic acid diarylamines andderivatives thereof. The disclosed diarylamines are pharmacologicallyactive.

BACKGROUND

MEK enzymes are dual specificity kinases involved in, for example,immunomodulation, inflammation, and proliferative diseases such ascancer and restenosis.

Proliferative diseases are caused by a defect in the intracellularsignaling system, or the signal transduction mechanism of certainproteins. Defects include a change either in the intrinsic activity orin the cellular concentration of one or more signaling proteins in thesignaling cascade. The cell may produce a growth factor that binds toits own receptors, resulting in an autocrine loop, which continuallystimulates proliferation. Mutations or overexpression of intracellularsignaling proteins can lead to spurious mitogenic signals within thecell. Some of the most common mutations occur in genes encoding theprotein known as Ras, a G-protein that is activated when bound to GTP,and inactivated when bound to GDP. The above-mentioned growth factorreceptors, and many other mitogenic receptors, when activated, lead toRas being converted from the GDP-bound state to the GTP-bound state.This signal is an absolute prerequisite for proliferation in most celltypes. Defects in this signaling system, especially in the deactivationof the Ras-GTP complex, are common in cancers, and lead to the signalingcascade below Ras being chronically activated.

Activated Ras leads in turn to the activation of a cascade ofserine/threonine kinases. One of the groups of kinases known to requirean active Ras-GTP for its own activation is the Raf family. These inturn activate MEK (e.g., MEK₁ and MEK₂) which then activates MAP kinase,ERK (ERK₁ and ERK₂). Activation of MAP kinase by mitogens appears to beessential for proliferation; constitutive activation of this kinase issufficient to induce cellular transformation. Blockade of downstream Rassignaling, for example by use of a dominant negative Raf-1 protein, cancompletely inhibit mitogenesis, whether induced from cell surfacereceptors or from oncogenic Ras mutants. Although Ras is not itself aprotein kinase, it participates in the activation of Raf and otherkinases, most likely through a phosphorylation mechanism. Onceactivated, Raf and other kinases phosphorylate MEK on two closelyadjacent serine residues, S²¹⁸ and S²²² in the case of MEK-1, which arethe prerequisite for activation of MEK as a kinase. MEK in turnphosphorylates MAP kinase on both a tyrosine, Y¹⁸⁵, and a threonineresidue, T¹⁸³, separated by a single amino acid. This doublephosphorylation activates MAP kinase at least 100-fold. Activated MAPkinase can then catalyze the phosphorylation of a large number ofproteins, including several transcription factors and other kinases.Many of these MAP kinase phosphorylations are mitogenically activatingfor the target protein, such as a kinase, a transcription factor, oranother cellular protein. In addition to Raf-1 and MEKK, other kinasesactivate MEK, and MEK itself appears to be a signal integrating kinase.Current understanding is that MEK is highly specific for thephosphorylation of MAP kinase. In fact, no substrate for MEK other thanthe MAP kinase, ERK, has been demonstrated to date and MEK does notphosphorylate peptides based on the MAP kinase phosphorylation sequence,or even phosphorylate denatured MAP kinase. MEK also appears toassociate strongly with MAP kinase prior to phosphorylating it,suggesting that phosphorylation of MAP kinase by MEK may require a priorstrong interaction between the two proteins. Both this requirement andthe unusual specificity of MEK are suggestive that it may have enoughdifference in its mechanism of action to other protein kinases thatselective inhibitors of MEK, possibly operating through allostericmechanisms rather than through the usual blockade of the ATP bindingsite, may be found.

SUMMARY

The invention features a compound having the formula (I) below:

R₁ is H, C₁₋₈ alkyl, C₃₋₈ alkenyl, C₃₋₈ alkynyl, C₃₋₈ cycloalkyl,phenyl, (phenyl)C₁₋₄ alkyl, (phenyl)C₃₋₄ alkenyl, (phenyl)C₃₋₄ alkynyl,(C₃₋₈ cycloalkyl)-C₁₋₄ alkyl, (C₃₋₈ cycloalkyl)C₃₋₄ alkenyl, (C₃₋₈cycloalkyl)C₃₋₄ alkynyl, C₃₋₈ heterocyclic radical, (C₃₋₈ heterocyclicradical)C₁₋₄ alkyl, (C₃₋₈ heterocyclic radical)C₃₋₄ alkenyl, (C₃₋₈heterocyclic radical)C₃₋₄ alkynyl, (CH₂)₂₋₄ OR_(C) or (CH₂)₂₋₄NR_(C)R_(D). R₂ is H, C₁₋₄ alkyl, phenyl, C₃₋₆ cycloalkyl, C₃₋₆heterocyclic radical, or (C₃₋₆ cycloalkyl)methyl. Each of R₃ and R₄ isindependently selected from H, F, NO₂, Br and Cl. R₅ is selected from Hand F. R₆ is H, F, Cl or CH₃. Each of R_(C) and R_(D) is independentlyselected from H, C₁₋₄ alkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, C₃₋₆cycloalkyl, and phenyl; or NR_(C)R_(D) may be a piperidino, morpholino,or N—(C₁₋₆ alkyl)piperazino ring. Each hydrocarbon radical above isoptionally substituted with between 1 and 3 substituents independentlyselected from halo, hydroxyl, amino, (amino)sulfonyl, and NO₂. Eachheterocyclic radical above is optionally substituted with between 1 and3 substituents independently selected from halo, C₁₋₄ alkyl, C₃₋₆cycloalkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, phenyl, hydroxyl, amino,(amino)sulfonyl, and NO₂, wherein each substituent alkyl, cycloalkyl,alkenyl, alkynyl or phenyl is in turn optionally substituted withbetween 1 and 3 substituents independently selected from halo, C₁₋₂alkyl, hydroxyl, amino, and NO₂. The invention also includes apharmaceutically acceptable salt or C₁₋₈ ester of a disclosed compound.For example, the disclosed alcohol compounds may form esters having thestructure obtained by replacing the H of a hydroxyl group with a—C(═O)C₁₋₇ acyl group.

The invention also relates to a pharmaceutical composition including (a)a compound of formula (I) and (b) a pharmaceutically-acceptable carrier.

The invention further relates to a method for treating proliferativediseases, such as cancer, restenosis, psoriasis, autoimmune disease, andatherosclerosis. Other aspects of the invention include methods fortreating MEK-related (including ras-related) cancers, whether solid orhematopoietic. Examples of cancers include colorectal, cervical, breast,ovarian, brain, acute leukemia, gastric, non-small cell lung,pancreatic, and renal cancer. Further aspects of the invention includemethods for treating or reducing the symptoms of xenograft (skin,cell(s), limb, organ or bone marrow transplant) rejection,osteoarthritis, rheumatoid arthritis, cystic fibrosis, complications ofdiabetes (including diabetic retinopathy and diabetic nephropathy),hepatomegaly, cardiomegaly, stroke (such as acute focal ischemic strokeand global cerebral ischemia), heart failure, septic shock, asthma, andAlzheimer's disease. Compounds of the invention are also useful asantiviral agents for treating viral infections such as HIV, hepatitis(B) virus (HBV), human papilloma virus (HPV), cytomegalovirus (CMV), andEpstein-Barr virus (EBV). These methods include the step ofadministering to a patient in need of such treatment, or suffering fromsuch a disease or condition, a pharmaceutically-effective amount of adisclosed compound or pharmaceutical composition thereof.

The invention also features methods of combination therapy, such as amethod for treating cancer, wherein the method further includesproviding radiation therapy or chemotherapy, for example, with mitoticinhibitors such as a taxane or a vinca alkaloid. Examples of mitoticinhibitors include paclitaxel, docetaxel, vincristine, vinblastine,vinorelbine, and vinflunine. Other therapeutic combinations include aMEK inhibitor of the invention and an anticancer agent such ascisplatin, 5-fluorouracil or 5-fluoro-2-4(1H,3H)-pyrimidinedione (5FU),flutamide, and gemcitabine.

The chemotherapy or radiation therapy may be administered before,concurrently, or after the administration of a disclosed compoundaccording to the needs of the patient.

The invention also includes synthetic intermediates and methodsdisclosed herein.

Other aspects of the invention are provided in the description,examples, and claims below.

DETAILED DESCRIPTION

The invention features diaryl amine compounds, pharmaceuticalcompositions thereof, and methods of using such compounds andcompositions.

According to one aspect of the invention, the compounds are MEKinhibitors. MEK inhibition assays include the in vitro cascade assay forinhibitors of MAP kinase pathway described at column 6, line 36 tocolumn 7, line 4 of U.S. Pat. No. 5,525,625 and the in vitro MEK assayat column 7, lines 4-27 of the same patent, the entire disclosure ofwhich is incorporated by reference (see also Examples 1-3 below). Awhole cell assay is described below in Example 4.

A. Terms

Certain terms are defined below and by their usage throughout thisdisclosure.

Alkyl groups include aliphatic (i.e., hydrocarbyl or hydrocarbon radicalstructures containing hydrogen and carbon atoms) with a free valence.Alkyl groups are understood to include straight chain and branchedstructures. Examples include methyl, ethyl, propyl, isopropyl, butyl,n-butyl, isobutyl, t-butyl, pentyl, isopentyl, 2,3-dimethylpropyl,hexyl, 2,3-dimethylhexyl, 1,1-dimethylpentyl, heptyl, and octyl.Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl.

Alkyl groups can be substituted with 1, 2, 3 or more substituents whichare independently selected from halo (fluoro, chloro, bromo, or iodo),hydroxy, amino, alkoxy, alkylamino, dialkylamino, cycloalkyl, aryl,aryloxy, arylalkyloxy, heterocyclic radical, and (heterocyclicradical)oxy. Specific examples include fluoromethyl, hydroxyethyl,2,3-dihydroxyethyl, (2- or 3-furanyl)methyl, cyclopropylmethyl,benzyloxyethyl, (3-pyridinyl)methyl, (2- or 3-furanyl)methyl,(2-thienyl)ethyl, hydroxypropyl, aminocyclohexyl, 2-dimethylaminobutyl,methoxymethyl, N-pyridinylethyl, diethylaminoethyl, andcyclobutylmethyl.

Alkenyl groups are analogous to alkyl groups, but have at least onedouble bond (two adjacent sp² carbon atoms). Depending on the placementof a double bond and substituents, if any, the geometry of the doublebond may be entgegen (E), or zusammen (Z), cis, or trans. Similarly,alkynyl groups have at least one triple bond (two adjacent sp carbonatoms). Unsaturated alkenyl or alkynyl groups may have one or moredouble or triple bonds, respectively, or a mixture thereof; like alkylgroups, unsaturated groups may be straight chain or branched, and theymay be substituted as described both above for alkyl groups andthroughout the disclosure by example. Examples of alkenyls, alkynyls,and substituted forms include cis-2-butenyl, trans-2-butenyl, 3-butynyl,3-phenyl-2-propynyl, 3-(2′-fluorophenyl)-2-propynyl,3-methyl(5-phenyl)-4-pentynyl, 2-hydroxy-2-propynyl,2-methyl-2-propynyl, 2-propenyl, 4-hydroxy-3-butynyl,3-(3-fluorophenyl)-2-propynyl, and 2-methyl-2-propenyl. In formula (I),alkenyl and alkynyl groups can be, for example, C₂₋₄, or C₂₋₈, but arepreferably C₃₋₄ or C₃₋₈.

More general forms of substituted hydrocarbon radicals includehydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl, hydroxycycloalkyl,hydroxyaryl, and corresponding forms for the prefixes amino-, halo-(e.g., fluoro-, chloro-, or bromo-), nitro-, alkyl-, phenyl-,cycloalkyl- and so on, or combinations of substituents. According toformula (I), therefore, substituted alkyls include hydroxyalkyl,aminoalkyl, nitroalkyl, haloalkyl, alkylalkyl (branched alkyls, such asmethylpentyl), (cycloalkyl)alkyl, phenylalkyl, alkoxy, alkylaminoalkyl,dialkylaminoalkyl, arylalkyl, aryloxyalkyl, arylalkyloxyalkyl,(heterocyclic radical)alkyl, and (heterocyclic radical)oxyalkyl. R₁ thusincludes hydroxyalkyl, hydroxyalkenyl, hydroxyalkynyl,hydroxycycloalkyl, hydroxyphenyl, hydroxy(phenyl)alkyl,(phenyl)hydryoxyalkyl, (C₃₋₈ hydroxylcycloalkyl)-C₁₋₄ alkyl, (C₃₋₈cycloalkyl)C₂₋₄ hydroxylalkenyl, C₃₋₈ hydroxyheterocyclic radical, (C₃₋₈heterocyclic radical)C₁₋₄ hydroxyalkyl, aminoalkyl, aminoalkenyl,aminoalkynyl, aminocycloalkyl, aminoaryl, alkylalkenyl,(alkylaryl)alkyl, (haloaryl)alkyl, (hydroxyaryl)alkynyl, and so forth.Similarly, R_(C) includes hydroxyalkyl and aminoaryl, and R_(D) includeshydroxyalkyl, aminoalkyl, and hydroxyalkyl(heterocyclic radical)alkyland so forth.

Heterocyclic radicals, which include but are not limited to heteroaryls,include: furyl, oxazolyl, isoxazolyl, thiophenyl, thiazolyl, pyrrolyl,imidazolyl, 1,3,4-triazolyl, tetrazolyl, pyridinyl, pyrimidinyl,pyridazinyl, indolyl, and their nonaromatic counterparts. Furtherexamples of heterocyclic radicals include piperidyl, quinolyl,isothiazolyl, piperidinyl, morpholinyl, piperazinyl, tetrahydrofuryl,tetrahydropyrrolyl, pyrrolidinyl, octahydroindolyl,octahydrobenzothiofuranyl, and octahydrobenzofuranyl.

Selective MEK 1 or MEK 2 inhibitors are those compounds which inhibitthe MEK 1 or MEK 2 enzymes, respectively, without substantiallyinhibiting other enzymes such as MKK3, PKC, Cdk2A, phosphorylase kinase,EGF, and PDGF receptor kinases, and C-src. In general, a selective MEK 1or MEK 2 inhibitor has an IC₅₀ for MEK 1 or MEK 2 that is at leastone-fiftieth ({fraction (1/50)}) that of its IC₅₀ for one of theabove-named other enzymes. Preferably, a selective inhibitor has an IC₅₀that is at least {fraction (1/100)}, more preferably {fraction (1/500)},and even more preferably {fraction (1/1000)}, {fraction (1/5000)}, orless than that of its IC₅₀ or one or more of the above-named enzymes.

B. Compounds

One aspect of the invention features the disclosed compounds shown informula (I) in the Summary section.

Embodiments of the invention include compounds wherein: (a) R₃ is bromoor chloro; (b) R₄ is fluoro; (c) R₅ is H; (d) each of R₄ and R₅ is H;(e) each of R₄ and R₅ is fluoro; (f) R₃ is bromo; (g) R₃ is fluoro; (h)R₄ is nitro; (i) R₅ is H; (j) R₆ is chloro; (k) R₆ is methyl; (l) R₁ isH or C₁₋₄ alkyl, and R₂ is H; (m) R₁ is (C₃₋₆ cycloalkyl)methyl; (n) R₁is H; (o) R₁ is (CH₂)₂₋₄OR_(C) or (CH₂)₂₋₄ NR_(C)R_(D); (p) R₆ is chloroor methyl; (q) R₆ is H; or combinations thereof.

Preferably, when R₁, R_(C), or R_(D) is an alkenyl or alkynyl, thedouble or triple bond, respectively, is not adjacent the point ofattachment when the point of attachment is a heteroatom. For example, R₁is preferably prop-2-ynyl, or but-2 or 3-enyl, and less preferablyprop-1-ynyl or but-1-enyl.

Examples of compounds of formula (I) include:4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonic acid;4-fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide;N-cyclopropylmethoxy-4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide;3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonic acid;3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide;N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide;3,4,5-trifluoro-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonic acid;3,4,5-trifluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide;N-cyclopropylmethoxy-3,4,5-trifluoro-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide;5-bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonicacid;5-bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide;5-bromo-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzenesulfonamide;2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzenesulfonic acid;N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzenesulfonamide; orN-cyclopropylmethoxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzenesulfonamide.

Further examples of compounds include:2-(2-chloro-4-iodo-phenylamino)-4-fluoro-benzenesulfonic acid;2-(2-chloro-4-iodo-phenylamino)-4-fluoro-N-hydroxy-benzenesulfonamide;2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-fluoro-benzenesulfonamide;2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-benzenesulfonic acid;2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzenesulfonamide;2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide;2-(2-chloro-4-iodo-phenylamino)-3,4,5-trifluoro-benzenesulfonic acid;2-(2-chloro-4-iodo-phenylamino)-3,4,5-trifluoro-N-hydroxy-benzenesulfonamide;2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-benzenesulfonamide;5-bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-benzenesulfonicacid;5-bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzenesulfonamide;5-bromo-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide;2-(2-chloro-4-iodo-phenylamino)-4-nitro-benzenesulfonic acid;2-(2-chloro-4-iodo-phenylamino)-N-hydroxy-4-nitro-benzenesulfonamide; or2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-nitro-benzenesulfonamide.

C. Synthesis

The disclosed compounds can be synthesized according to Scheme 1 below.

One equivalent of appropriately substituted sulfonyl chloride is addedto a solution of one equivalent of appropriately substitutedhydroxylamine and excess triethylamine in CH₂Cl₂ or Et₂O and stirred for30 minutes. The triethylamine hydrochloride precipitate is separated byfiltration and discarded. If necessary, the product is further purifiedby chromatography on silica column. The pure 2-fluor hydroxamic orhydroxamate product is then added to a solution of appropriatelysubstituted lithium anilide prepared by adding LDA to the aniline in THFat −78° C. After stirring at room temperature for 16 hours, the reactionmixture is poured in to Et₂O—HCl. Any precipitated solid is separated byfiltration and discarded. The filtrate is concentrated and the resultingcrude product is purified on silica column to give the desired targetproduct.

The disclosed compounds can also be made by other synthetic organicmethods, as well as automated or combinatorial methods.

D. Uses

The disclosed compositions are useful as both prophylactic andtherapeutic treatments for diseases or conditions as provided in theSummary section, as well as diseases or conditions modulated by the MEKcascade. Examples include stroke, heart failure, osteoarthritis,rheumatoid arthritis, organ transplant rejection, and a variety oftumors such as ovarian, lung, pancreatic, brain, prostatic, andcolorectal.

1. Dosages

Those skilled in the art will be able to determine, according to knownmethods, the appropriate dosage for a patient, taking into accountfactors such as age, weight, general health, the type of pain requiringtreatment, and the presence of other medications. In general, aneffective amount will be between 0.1 and 1000 mg/kg per day, preferablybetween 1 and 300 mg/kg body weight, and daily dosages will be between10 and 5000 mg for an adult subject of normal weight. Commerciallyavailable capsules or other formulations (such as liquids andfilm-coated tablets) of 100 mg, 200 mg, 300 mg, or 400 mg can beadministered according to the disclosed methods.

2. Formulations

Dosage unit forms include tablets, capsules, pills, powders, granules,aqueous and nonaqueous oral solutions and suspensions, and parenteralsolutions packaged in containers adapted for subdivision into individualdoses. Dosage unit forms can also be adapted for various methods ofadministration, including controlled release formulations, such assubcutaneous implants. Administration methods include oral, rectal,parenteral (intravenous, intramuscular, subcutaneous), intracisternal,intravaginal, intraperitoneal, intravesical, local (drops, powders,ointments, gels, or cream), and by inhalation (a buccal or nasal spray).

Parenteral formulations include pharmaceutically acceptable aqueous ornonaqueous solutions, dispersion, suspensions, emulsions, and sterilepowders for the preparation thereof. Examples of carriers include water,ethanol, polyols (propylene glycol, polyethylene glycol), vegetableoils, and injectable organic esters such as ethyl oleate. Fluidity canbe maintained by the use of a coating such as lecithin, a surfactant, ormaintaining appropriate particle size. Carriers for solid dosage formsinclude (a) fillers or extenders, (b) binders, (c) humectants, (d)disintegrating agents, (e) solution retarders, (f) absorptionaccelerators, (g) adsorbants, (h) lubricants, (i) buffering agents, and(j) propellants.

Compositions may also contain adjuvants such as preserving, wetting,emulsifying, and dispensing agents; antimicrobial agents such asparabens, chlorobutanol, phenol, and sorbic acid; isotonic agents suchas a sugar or sodium chloride; absorption-prolonging agents such asaluminum monostearate and gelatin; and absorption-enhancing agents.

3. Related Compounds

The invention provides the disclosed compounds and closely related,pharmaceutically acceptable forms of the disclosed compounds, such assalts, esters, amides, hydrates or solvated forms thereof; masked orprotected forms; and racemic mixtures, or enantiomerically or opticallypure forms.

Pharmaceutically acceptable salts, esters, and amides includecarboxylate salts (e.g., C₁₋₈ alkyl, cycloalkyl, aryl, heteroaryl, ornon-aromatic heterocyclic), amino acid addition salts, esters, andamides which are within a reasonable benefit/risk ratio,pharmacologically effective, and suitable for contact with the tissuesof patients without undue toxicity, irritation, or allergic response.Representative salts include hydrobromide, hydrochloride, sulfate,bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate,stearate, laurate, borate, benzoate, lactate, phosphate, tosylate,citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate,glucoheptonate, lactiobionate, and laurylsulfonate. These may includealkali metal and alkali earth cations such as sodium, potassium,calcium, and magnesium, as well as non-toxic ammonium, quaternaryammonium, and amine cations such as tetramethyl ammonium, methylamine,trimethylamine, and ethylamine. See, for example, S. M. Berge, et al.,“Pharmaceutical Salts,” J. Pharm. Sci., 1977, 66:1-19 which isincorporated herein by reference. Representative pharmaceuticallyacceptable amides of the invention include those derived from ammonia,primary C₁₋₆ alkyl amines and secondary di (C₁₋₆ alkyl)amines. Secondaryamines include 5- or 6-membered heterocyclic or heteroaromatic ringmoieties containing at least one nitrogen atom and optionally between 1and 2 additional heteroatoms. Preferred amides are derived from ammonia,C₁₋₃ alkyl primary amines, and di (C₁₋₂ alkyl)amines. Representativepharmaceutically acceptable esters of the invention include C₁₋₇ alkyl,C₅₋₇ cycloalkyl, phenyl, and phenyl(C₁₋₆)alkyl esters. Preferred estersinclude methyl esters.

The invention also includes disclosed compounds having one or morefunctional groups (e.g., hydroxyl, amino, or carboxyl) masked by aprotecting group. Some of these masked or protected compounds arepharmaceutically acceptable; others will be useful as intermediates.Synthetic intermediates and processes disclosed herein, and minormodifications thereof, are also within the scope of the invention.

Hydroxyl Protecting Groups

Hydroxyl protecting groups include: ethers, esters, and protection for1,2- and 1,3-diols. The ether protecting groups include: methyl,substituted methyl ethers, substituted ethyl ethers, substituted benzylethers, silyl ethers and conversion of silyl ethers to other functionalgroups.

Substituted Methyl Ethers

Substituted methyl ethers include: methoxymethyl, methylthiomethyl,t-utylthiomethyl, (phenyldimethylsilyl)methoxymethyl, benzyloxymethyl,p-ethoxybenzyloxymethyl, (4-methoxyphenoxy)methyl, guaiacolmethyl,t-butoxymethyl, 4-pentenyloxymethyl, siloxymethyl,2-methoxyethoxymethyl, 2,2,2-trichloroethoxymethyl, bis(2-chloro-ethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl, tetrahydropyranyl,3-bromotetrahydro-pyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl,4-methoxytetrahydropyranyl, 4-methoxytetrahydrothiopyranyl,4-methoxytetrahydrothiopyranyl S,S-dioxido,1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl, 1,4-dioxan-2-yl,tetrahydrofuranyl, tetrahydrothiofuranyl, and2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-ethanobenzofuran-2-yl.

Substituted Ethyl Ethers

Substituted ethyl ethers include: 1-ethoxyethyl,1-(2,chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilyethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, andbenzyl.

Substituted Benzyl Ethers

Substituted benzyl ethers include: p-methoxybenzyl, 3,4-dimethoxybenzyl,o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl,p-cyanobenzyl, p-phenylbenzyl, 2- and 4-picolyl, 3-methyl-2-picolylN-oxido, diphenylmethyl, p, p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, α-naphthyldiphenyl-methyl,p-methoxyphenyldiphenylmethyl, di (p-methoxyphenyl)phenylmethyl,tri-(p-methoxyphenyl)methyl,4-(4′-bromophenacyloxy)phenyldiphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimido-phenyl)methyl,4,4′,4″-tris(levulinoyloxy-phenyl)methyl,4,4′,4″tris(benzoyloxy-phenyl)methyl,3-(imidazol-1-ylmethyl)-bis(4′,4″-dimethoxyphenyl)-methyl,1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodithiolan-2-yl, and benzisothiazolyl S,S-dioxido.

Silyl Ethers

Silyl ethers include: trimethylsilyl, triethylsilyl, triisopropylsilyl,dimethylisopropylsilyl, diethylisopropylsilyl, dimethylthexylsilyl,t-butyidimethylsilyl, t-butyldiphenylsilyl, tribenzylsilyl,tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl, andt-butylmethoxy-phenylsilyl.

Esters

Esters protecting groups include: esters, carbonates, assisted cleavage,miscellaneous esters, and sulfonates.

Esters

Examples of protective esters include: formate, benzoylformate, acetate,chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate,methoxyacetate, triphenylmethoxyacetate, phenoxyacetate,p-chlorophenoxyacetate, p-P-phenylacetate, 3-phenylpropionate,4-oxopentanoate(levulinate), 4,4-(ethylenedithio)pentanoate, pivaloate,adamantoate,crotonate,4-methoxycrotonate, benzoate, p-phenylbenzoate,and 2,4,6-trimethylbenzoate(mesitoate).

Carbonates

Carbonates include: methyl, 9-fluorenylmethyl, ethyl,2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl,2-(triphenylphosphonio)ethyl, isobutyl, vinyl, allyl, p-nitrophenyl,benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, S-benzyl thiocarbonate, 4-ethoxy-1-naphthyl, and methyldithiocarbonate.

Assisted Cleavage

Examples of assisted cleavage protecting groups include: 2-iodobenzoate,4-azido-butyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzene-sulfonate, 2-(methylthiomethoxy)ethyl carbonate,4-(methylthiomethoxymethyl)benzoate, and2-(methylthiomethoxymethyl)benzoate.

Miscellaneous Esters

In addition to the above classes, miscellaneous esters include:2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate (tigloate),o-(methoxycarbonyl) benzoate, p-P-benzoate, α-naphthoate, nitrate, alkylN,N,N′N′-tetramethyl-phosphorodiamidate, N-phenylcarbamate, borate,dimethylphosphinothioyl, and 2,4-dinitrophenylsulfenate.

Sulfonates

Protective sulfates includes: sulfate, methanesulfonate(mesylate),benzylsulfonate, and tosylate.

Protection for 1,2- and 1,3-diols

The protection for 1,2 and 1,3-diols group includes: cyclic acetals andketals, cyclic ortho esters, and silyl derivatives.

Cyclic Acetals and Ketals

Cyclic acetals and ketals include: methylene, ethylidene,1-t-butylethylidene, 1-phenylethylidene, (4-methoxyphenyl)ethylidene,2,2,2-trichloroethylidene, acetonide (isopropylidene), cyclopentylidene,cyclohexylidene, cycloheptylidene, benzylidene, p-methoxybenzylidene,2,4-dimethoxybenzylidene, 3,4-dimethoxybenzylidene, and2-nitrobenzylidene.

Cyclic Ortho Esters

Cyclic ortho esters include: methoxymethylene, ethoxymethylene,dimethoxy-methylene, 1-methoxyethylidene, 1-ethoxyethylidine,1,2-dimethoxyethylidene, α-methoxybenzylidene,1-(N,N-dimethylamino)ethylidene derivative,α-(N,N-dimethylamino)benzylidene derivative, and 2-oxacyclopentylidene.

Protection for the Carboxyl Group Esters

Ester protecting groups include: esters, substituted methyl esters,2-substituted ethyl esters, substituted benzyl esters, silyl esters,activated esters, miscellaneous derivatives, and stannyl esters.

Substituted Methyl Esters

Substituted methyl esters include: 9-fluorenylmethyl, methoxymethyl,methylthiomethyl, tetrahydropyranyl, tetrahydrofuranyl,methoxyethoxymethyl, 2-(trimethylsilyl)ethoxy-methyl, benzyloxymethyl,phenacyl, p-bromophenacyl, α-methylphenacyl, p-methoxyphenacyl,carboxamidomethyl, and N-phthalimidomethyl.

2-Substituted Ethyl Esters

2-Substituted ethyl esters include: 2,2,2-trichloroethyl, 2-haloethyl,|-chloroalkyl, 2-(trimethylsily)ethyl, 2-methylthioethyl,1,3-dithianyl-2-methyl, 2(p-nitrophenylsulfenyl)-ethyl,2-(p-toluenesulfonyl)ethyl, 2-(2′-pyridyl)ethyl,2-(diphenylphosphino)ethyl, 1-methyl-1-phenylethyl, t-butyl,cyclopentyl, cyclohexyl, allyl, 3-buten-1-yl,4-(trimethylsily)-2-buten-1-yl, cinnamyl, α-methylcinnamyl, phenyl,p-(methylmercapto)-phenyl, and benzyl.

Substituted Benzyl Esters

Substituted benzyl esters include: triphenylmethyl, diphenylmethyl,bis(o-nitrophenyl)methyl, 9-anthrylmethyl, 2-(9,10-dioxo)anthrylmethyl,5-dibenzo-suberyl, 1-pyrenylmethyl ,2-(trifluoromethyl)-6-chromylmethyl,2,4,6-trimethyl-benzyl, p-bromobenzyl, o-nitrobenzyl, p-nitrobenzyl,p-methoxybenzyl, 2,6-dimethoxybenzyl, 4-(methylsulfinyl)benzyl,4-sulfobenzyl, piperonyl, and 4-P-benzyl.

Silyl Esters

Silyl esters include: trimethylsilyl, triethylsilyl,t-butyldimethylsilyl, i-propyldimethylsilyl, phenyidimethylsilyl, anddi- t-butylmethylsilyl.

Miscellaneous Derivatives

Miscellaneous derivatives includes: oxazoles, 2-alkyl-1,3-oxazolines,4-alkyl-5-oxo-1,3-oxazolidines, 5-alkyl-4-oxo-1,3-dioxolanes, orthoesters, phenyl group, and pentaaminocobalt(III) complex.

Stannyl Esters

Examples of stannyl esters include: triethylstannyl andtri-n-butylstannyl.

Amides and Hydrazides

Amides include: N,N-dimethyl, pyrrolidinyl, piperidinyl,5,6-dihydrophen-anthridinyl, o-nitroanilides, N-7-nitroindolyl,N-8-nitro-1,2,3,4-tetrahydroquinolyl, and p-P-benzenesulfonamides.Hydrazides include: N-phenyl, N,N′-diisopropyl and other dialkylhydrazides.

Protection for the Amino Group

Carbamates

Carbamates include: carbamates, substituted ethyl, assisted cleavage,photolytic cleavage, urea-type derivatives, and miscellaneouscarbamates.

Carbamates

Carbamates include: methyl and ethyl, 9-fluorenylmethyl,9-(2-sulfo)fluorenylmethyl, 9-(2,7-dibromo)fluorenylmethyl,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydro-thioxanthyl)]methyl, and4-methoxyphenacyl.

Substituted Ethyl

Substituted ethyl protective groups include: 2,2,2-trichloroethyl,2-trimethylsilylethyl, 2-phenylethyl, 1-(1-adamantyl)-1-methylethyl,1,1-dimethyl-2-haloethyl, 1,1dimethyl-2,2-dibromoethyl,1,1-dimethyl-2,2,2-trichloroethyl, 1-methyl-1-(4-biphenylyl)ethyl,1-(3,5-di-t-butylphenyl)-1-methylethyl, 2-(2′-and 4′-pyridyl)ethyl,2-(N,N-icyclohexylcarboxamido)- ethyl, t-butyl, 1-adamantyl, vinyl,allyl, 1-isopropylallyl, connamyl, 4-nitrocinnamyl, quinolyl,N-hydroxypiperidinyl, alkyldithio, benzyl, p-methoxybenzyl,p-nitrobenzyl, p-bromobenzyl, p-chlorobenzyl, 2,4dichlorobenzyl,4-methylsulfinylbenzyl, 9-anthrylmethyl, and diphenylmethyl.

Assisted Cleavage

Protection via assisted cleavage includes: 2-methylthioethyl,2-methylsulfonylethyl, 2-(p-toluenesulfonyl)ethyl,[2-(1,3-dithianyl)]methyl, 4-methylthiophenyl, 2,4-dimethyl-thiophenyl,2-phosphonioethyl, 2-triphenylphosphonioisopropyl,1,1-dimethyl-2cyanoethyl, m-chloro-p-acyloxybenzyl,p-(dihydroxyboryl)benzyl, 5-benzisoxazolyl-methyl, and2-(trifluoromethyl)-6-chromonylmethyl.

Photolytic Cleavage

Photolytic cleavage methods use groups such as: m-nitrophenyl,3,5-dimethoxybenzyl, o-nitrobenzyl, 3,4-dimethoxy-6-nitrobenzyl, andphenyl(o-nitrophenyl)methyl.

Urea-Type Derivatives

Examples of of urea-type derivatives include:phenothiazinyl-(10)-carbonyl derivative,N′-p-toluenesulfonylaminocarbonyl, and N′-phenylaminothiocarbonyl.

Miscellaneous Carbamates

In addition to the above, miscellaneous carbamates include: t-amyl,S-benzyl thiocarbamate, p-cyanobenzyl, cyclobutyl, cyclohexyl,cyclopentyl, cyclopropylmethyl, p-decyloxy-benzyl, diisopropylmethyl,2,2-dimethoxy-carbonylvinyl, o-(N,N-dimethyl-carboxamido)-benzyl,1,1-dimethyl-3(N,N-dimethylcarboxamido)propyl, 1,1-dimethyl-propynyl,di(2-pyridyl)methyl, 2-furanylmethyl, 2-iodoethyl, isobornyl, isobutyl,isonicotinyl, p(p′-methoxyphenyl-azo)benzyl, 1-methylcyclobutyl,1-methylcyclohexyl, 1-methyl-1-cyclopropyl-methyl,1-methyl-(3,5-dimethoxyphenyl)ethyl, 1-methyl-1(p-henylazophenyl)-ethyl,1-methyl-1-phenylethyl, 1-methyl-1-(4-pyridyl)ethyl, phenyl,p-(phenylazo)benzyl, 2,4,6-tri-t-butylphenyl,4-(trimethylammonium)benzyl, and 2,4,6-trimethylbenzyl.

Amides

Amides

Amides includes: N-formyl, N-acetyl, N-chloroacetyl, N-trichloroacetyl,N-trifluoroacetyl, N-phenylacetyl, N-3-phenylpropionyl, N-picolinoyl,N-3-pyridyl-carboxamide, N-benzoylphenylalanyl derivative, N-benzoyl,and N-p-phenylbenzoyl.

Assisted Cleavage

Assisted cleavage groups include: N-o-nitrophenylacetyl,N-o-nitrophenoxyacetyl, N-acetoacetyl,(N′-dithiobenzyloxycarbonylamino)acetyl, N-3-(p-hydroxphenyl)propionyl,N-3-(o-nitrophenyl)propionyl, N-2-methyl-2-(o-nitrophenoxy)propionyl,N-2-methyl-2-(o-phenylazophenoxy)propionyl, N-4-chlorobutyryl,N-3-methyl-3-nitrobutyryl, N-o-nitrocinnamoyl, N-acetylmethioninederivative, N-o-nitrobenzoyl, N-o-(benzoyloxymethyl)benzoyl, and4,5-diphenyl-3-oxazolin-2-one.

Cyclic Imide Derivatives

Cyclic imide derivatives include: N-phthalimide, N-dithiasuccinoyl,N-2,3-diphenyl-maleoyl, N-2,5-dimethylpyrrolyl,N-1,1,4,4-tetramethyl-disilylazacyclopentane adduct, 5-substituted1,3-dimethyl-1,3,5-triaza-cyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, and 1-substituted3,5-dinitro-4-pyridonyl.

Special —NH Protective Groups

Protective groups for —NH include: N-alkyl and N-aryl amines, iminederivatives, enamine derivatives, and N-hetero atom derivatives (such asN-metal, N—N, N—P, N—Si, and N—S), N-sulfenyl, and N-sulfonyl.

N-Alkyl and N-Aryl Amines

N-alkyl and N-aryl amines include: N-methyl, N-allyl,N-[2-(trimethylsilyl)ethoxyl]-methyl, N-3-acetoxypropyl,N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl), quaternary ammoniumsalts, N-benzyl, N-di(4-methoxyphenyl)methyl, N-5-dibenzosuberyl,N-triphenylmethyl, N-(4-methoxyphenyl)diphenylmethyl,N-9-phenylfluorenyl, N-2,7-dichloro-9-fluorenylmethylene,N-ferrocenylmethyl, and N-2-picolylamine N′-oxide.

Imine Derivatives

Imine derivatives include: N-1,1-dimethylthiomethylene, N-benzylidene,N-p-methoxybenzylidene, N-diphenylmethylene,N-[(2-pyridyl)mesityl]methylene, N-(N′,N′-dimethylaminomethylene), N,N′-isopropylidene, N-p-nitrobenzylidene, N-salicylidene,N-5-chlorosalicylidene, N-(5-chloro-2-hydroxyphenyl)phenyl-methylene,and N-cyclohexylidene.

Enamine Derivative

An example of an enamine derivative isN-(5,5-dimethyl-3-oxo-1-cyclohexenyl).

N-Hetero Atom Derivatives

N-metal derivatives include: N-borane derivatives, N-diphenylborinicacid derivative, N-[phenyl (pentacarbonylchromium- or-tungsten)]carbenyl, and N-copper or N-zinc chelate. Examples of N—Nderivatives include: N-nitro, N-nitroso, and N-oxide. Examples of N—Pderivatives include: N-diphenylphosphinyl, N-dimethylthiophosphinyl,N-diphenylthiophosphinyl, N-dialkyl phosphoryl, N-dibenzyl phosphoryl,and N-diphenyl phosphoryl. Examples of N-sulfenyl derivatives include:N-benzenesulfenyl, N-o-nitrobenzenesulfenyl,N-2,4-dinitrobenzenesulfenyl, N-pentachlorobenzenesulfenyl,N-2-nitro-4-methoxy-benzenesulfenyl, N-triphenylmethylsulfenyl, andN-3-nitropyridinesulfenyl. N-sulfonyl derivatives include:N-p-toluenesulfonyl, N-benzenesulfonyl,N-2,3,6-trimethyl-4-methoxybenzenesulfonyl,N-2,4,6-trimethoxybenzenesulfonyl,N-2,6-dimethyl-4-methoxy-benzenesulfonyl, N-pentamethylbenzenesulfonyl,N-2,3,5, 6-tetramethyl-4-methoxybenzene-sulfonyl,N-4-methoxybenzenesulfonyl, N-2,4,6-trimethylbenzenesulfonyl,N-2,6-dimethoxy-4-methylbenzenesulfonyl,N-2,2,5,7,8-pentamethylchroman-6-sulfonyl, N-methanesulfonyl,N-β-trimethylsilylethanesulfonyl, N-9-anthracenesulfonyl,N-4-(4′,8′-dimethoxynaphthylmethyl)-benzenesulfonyl, N-benzylsulfonyl,N-trifluoromethylsulfonyl, and N-phenacylsulfonyl.

Disclosed compounds which are masked or protected may be prodrugs,compounds metabolized or otherwise transformed in vivo to yield adisclosed compound, e.g., transiently during metabolism. Thistransformation may be a hydrolysis or oxidation which results fromcontact with a bodily fluid such as blood, or the action of acids, orliver, gastrointestinal, or other enzymes.

Features of the invention are further described in the examples below.

EXAMPLES Synthetic Example Preparation of2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide(PD 0297447)

N-cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide.

To a stirring suspension comprised of O-cyclopropylmethyl-hydroxylaminehydrochloride (5.40 g, 0.0437 mol) in dichloromethane (20 ml) at ambienttemperature under a nitrogen atmosphere was added diisopropylethylamine(10.8 ml, 0.062 mol). A solution comprised of2,3,4-trifluorobenzenesulfonyl chloride (Oakwood Products, Inc., 1.00 g,4.34×10⁻³ mol) in dichloromethane (120 ml) was added dropwise to thereaction vessel containing the stirring suspension over a 12 minuteperiod. The reaction mixture was stirred for another 12 minutes and wasquenched with 10% aqueous hydrochloric acid (140 ml). The biphasicmixture was stirred vigorously for 16 hours. The layers were separatedand the organic phase was dried (MgSO₄) and concentrated to 6 ml volume.The concentrated solution was administered to a flash silica column(Biotage, 90 g of silica gel). Elution with dichloromethane afforded0.8283 g of a white amorphous solid; 68% yield; ¹H-NMR (400 MHz; CDCl₃signal offset to δ 7.03; values reported are uncorrected) δ 7.50 (m,1H),7.10 (s,1H), 6.95 (m,1H), 3.59 (d, 2H, J═7.2 Hz), 0.80 (m, 1H), 0.31 (m,2H), 0.02 (m, 2H); ¹⁹F-NMR (376 MHz; CDCl₃) δ−122.65 (m, 1F), −129.37(m, 1F), −156.20 (m, 1F); MS (APCl−) 280 (M−1, 100), 210 (55), 195 (45).

2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzenesulfonamide(PD 0297447).

To a stirring solution comprised of 2-chloro-4-iodoaniline intetrahydrofuran (10 ml) at −78° C. under a nitrogen atmosphere was addeda 1.0 M tetrahydrofuran solution of lithium bistrimethylsilylamide (6.2ml, 6.2×10⁻³ mol) to form a green suspension. The suspension was stirredfor five minutes before a stirring suspension comprised of lithiatedN-cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide (prepared byadding 3.0 ml of the 1.0 M lithium bistrimethylsilylamide solution to astirring solution comprised ofN-cyclopropylmethoxy-2,3,4-trifluoro-benzenesulfonamide in 10 ml oftetrahydrofuran at −78° C. under nitrogen gas) was added via canula. Thecold bath was removed and the stirring suspension was stirred for onehour. The reaction mixture was quenched with 10% aqueous hydrochloricacid (50 ml) and the biphasic mixture was concentrated in vacuo to anaqueous suspension that was extracted with diethyl ether (200 ml). Theorganic phase was dried (MgSO₄) an d was concentrated in vacuo to afforda tan oil. The crude product was purified by flash chromatography.Elution with a gradient (hexanes-ethyl acetate 99:1→(2 min) 9:1→(25 min)3:1 afforded 1.10 g of a white amorphous foam; 73% yield; ¹H-NMR (400MHz; DMSO) δ 7.69 (m, 1H), 7.59 (d, 1H, J=1.9 Hz), 7.34 (dd, 1H, J=8.7,1.9 Hz), 7.27 (s, 1H), 7.00 (s, 1H), 6.95 (m, 1H), 6.43 (dd, 1H, J=8.7,5.8 Hz), 3.52 (d, 2H, J=7.5 Hz), 0.74 (m, 1H), 0.34 (m, 2H), 0.02 (m,2H); ¹⁹F-NMR (376 MHz; CDCl₃) δ−124.76 (m, 1F), −136.69 (d, 1F, J=18.3Hz); MS (APCl+) 515 (M+1, 100); (APCl−) 513 (M−1, 50), 443 (73), 428(100); IR (KBr) 1491 cm⁻¹; Anal. Calcd/found for C₁₆H₁₄CIF₂IN₂O₃S C,37.34/36.54; H, 2.74/2.71; N, 5.44/5.15; F, 7.38/7.57.

The APK IC₅₀ for PD 0297447 is 0.965 μM.

Biological Examples Example 1

Cascade Assay for Inhibitors of the MAP Kinase Pathway

Incorporation of ³²P into myelin basic protein (MBP) is assayed in thepresence of a glutathione S-transferase fusion protein containing p44MAPkinase (GST-MAPK) and a glutathione S-transferase fusion proteincontaining p45MEK (GST-MEK). The assay solution contains 20 mM HEPES, pH7.4, 10 mM MgCl₂, 1 mM MnCl₂, 1 mM EGTA, 50,μM [γ³²P]ATP, 10 μg GST-MEK,0.5 μg GST-MAPK and 40 μg MBP in a final volume of 100 μL. Reactions arestopped after 20 minutes by addition of trichloroacetic acid andfiltered through a GF/C filter mat. ³²P retained on the filter mat isdetermined using a 120S Betaplate. Compounds are assessed at 10 μM forability to inhibit incorporation of 32P.

To ascertain whether compounds are inhibiting GST-MEK or GST MAPK, twoadditional protocols are employed. In the first protocol, compounds areadded to tubes containing GST-MEK, followed by addition of GST-MAPK, MBPand [γ-³²P]ATP. In the second protocol, compounds are added to tubescontaining both GST-MEK and GST-MAPK, followed by MBP and [γ³²P]ATP.

Compounds that show activity in both protocols are scored as MAPKinhibitors, while compounds showing activity in only the first protocolare scored as MEK inhibitors.

Example 2

In Vitro MAP Kinase Assay

Inhibitory activity can be confirmed in direct assays. For MAP kinase, 1μg GST-MAPK is incubated with 40 μg MBP for 15 minutes at 30° C. in afinal volume of 50 μL containing 50 mM Tris (pH 7.5), 10 μM MgC1₂, 2 μMEGTA, and 10 μM [γ-³²P]ATP. The reaction is stopped by addition ofLaemmli SDS sample buffer and phosphorylated MBP resolved byelectrophoresis on a 10% polyacrylamide gel. Radioactivity incorporatedinto MBP is determined by both autoradiography, and scintillationcounting of excised bands.

Example 3

In Vitro MEK Assay

For evaluation of direct MEK activity, 10 μg GST-MEK₁ is incubated with5 μg of a glutathione S-transferase fusion protein containing p44MAPkinase with a lysine to alanine mutation at position 71 (GST-MAPK-KA).This mutation eliminates kinase activity of MAPK, so only kinaseactivity attributed to the added MEK remains. Incubations are 15 minutesat 30° C. in a final volume of 50 μL containing 50 mM Tris (pH 7.5), 10μM MgCl₂, 2, μM EGTA, and 10 μM [γ-³²P]ATP. The reaction is stopped byaddition of Laemrnli SDS sample buffer. Phosphorylated GST-MAPK-KA isresolved by electrophoresis on a 10% polyacrylamide gel. Radioactivityincorporated into GST-MAPK-KA is determined by autoradiography, andsubsequent scintillation counting of excised bands. Additionally, anartificially activated MEK containing serine to glutamate mutations atpositions 218 and 222 (GST-MEK-2E) is used. When these two sites arephosphorylated, MEK activity is increased. Phosphorylation of thesesites can be mimicked by mutation of the serine residues to glutamate.For this assay, 5 μg GST-MEK-2E is incubated with 5 μg GST-MAPK-KA for15 minutes at 30° C. in the same reaction buffer as described above.Reactions are terminated and analyzed as above.

Example 4

Whole Cell MAP Kinase Assay

To determine if compounds block activation of MAP kinase in whole cells,the following protocol is used. Cells are plated in multi-well platesand grown to confluence. Cells are serum-deprived overnight. Cells areexposed to the desired concentrations of compound or vehicle (DMSO) for30 minutes, followed by addition of a growth factor, for example, PDGF(100 ng/mL). After a 5-minute treatment with the growth factor, cellsare washed with PBS, and lysed in a buffer consisting of 70 mM NaCl, 10mM HEPES (pH 7.4), 50 mM glycerol phosphate, and 1% Triton X-1 00.Lysates are clarified by centrifugation at 13,000×g for 10 minutes. Fivemicrograms of the resulting supernatants are incubated with 10 μgmicrotubule associated protein-2 (Map2) for 15 minutes at 30° C. in afinal volume of 25 μL containing 50 mM Tris (pH 7.4), 10 mM MgCl₂, 2 mMEGTA and 30 μM [γ-³²P]ATP. Reactions are terminated by addition ofLaermmli sample buffer. Phosphorylated Map2 is resolved on 7.5%acrylamide gels and incorporated radioactivity is determined byscintillation counting of excised bands.

Example 5

Monolayer Growth

Cells are plated into multi-well plates at 10 to 20,000 cells/mL.Forty-eight hours after seeding, test compounds are added to the cellgrowth medium and incubation is continued for 2 additional days. Cellsare then removed from the wells by incubation with trypsin andenumerated with a Coulter counter.

Example 6

Growth in Soft-Agar

Cells are seeded into 35-mm dishes at 5 to 10,000 cells/dish usinggrowth medium containing 0.3% agar. After chilling to solidify the agar,cells are transferred to a 37° C. incubator. After 7 to 10 days' growth.visible colonies are manually enumerated with the aid of a dissectingmicroscope.

Example 7

Collagen-Induced Arthritis in Mice

Type II collagen-induced arthritis (CIA) in mice is an experimentalmodel of arthritis that has a number of pathologic, immunologic, andgenetic features in common with rheumatoid arthritis. The disease isinduced by immunization of DBA/1 mice with 100 μg type II collagen,which is a major component of joint cartilage, delivered intradermallyin Freund's complete adjuvant. The disease susceptibility is regulatedby the class II MHC gene locus, which is analogous to the association ofrheumatoid arthritis with HLA-DR4.

A progressive and inflammatory arthritis develops in the majority ofmice immunized, characterized by paw width increases of up to 100%. Atest compound is administered to mice in a range of amounts, such as 20,60, 100, and 200 mg/kg body weight/day. The duration of the test can beseveral weeks to a few months, such as 40, 60, or 80 days. A clinicalscoring index is used to assess disease progression from erythema andedema (stage 1), joint distortion (stage 2), to joint ankylosis (stage3). The disease is variable in that it can affect one or all paws in ananimal, resulting in a total possible score of 12 for each mouse.Histopathology of an arthritic joint reveals synovitis, pannusformation, and cartilage and bone erosions. All mouse strains that aresusceptible to CIA are high antibody responders to type II collagen, andthere is a marked cellular response to CII.

Example 8

SCW-Induced Monoarticular Arthritis

Arthritis is induced as described by Schwab, et al., Infection andImmunity, 59:4436-4442 (1991) with minor modifications. Rats receive 6μg sonicated SCW [in 10 μl Dulbecco's PBS (DPBS)] by an intraarticularinjection into the right tibiotalar joint on day 0. On day 21, the DTHis initiated with 100 μg of SCW (250 μl) administered i.v. For oralcompound studies, compounds are suspended in vehicle (0.5%hydroxypropyl-methylcellulose/0.2% Tween 80), sonicated, andadministered twice daily (10 ml/kg volume) beginning 1 hr prior toreactivation with SCW. Compounds are administered in amounts between 10and 500 mg/kg body weight/day, such as 20, 30, 60,100, 200, and 300mg/kg/day. Edema measurements are obtained by determining the baselinevolumes of the sensitized hindpaw before reactivation on day 21, andcomparing them with volumes at subsequent time points such as day 22,23, 24, and 25. Paw volume is determined by mercury plethysmography.

Example 9

Mouse Ear-Heart Transplant Model

Fey, T. A. et al. describe methods for transplanting split-heartneonatal cardiac grafts into the ear pinna of mice and rats (J. Pharm.and Toxic. Meth. 39:9-17 (1998)). Compounds are dissolved in solutionscontaining combinations of absolute ethanol, 0.2% hydroxypropylmethylcellulose in water, propylene glycol, cremophor, and dextrose, orother solvent or suspending vehicle. Mice are dosed orally orintraperitoneally once, twice or three times daily from the day oftransplant (day 0) through day 13 or until grafts have been rejected.Rats are dosed once, twice, or three times daily from day 0 through day13. Each animal is anesthetized and an incision is made at the base ofthe recipient ear, cutting only the dorsal epidermis and dermis. Theincision is spread open and down to the cartilage parallel to the head,and sufficiently wide to accommodate the appropriate tunneling for a rator insertion tool for a mouse. A neonatal mouse or rat pup less than 60hours old is anesthetized and cervically dislocated. The heart isremoved from the chest, rinsed with saline, bisected longitudinally witha scalpel, and rinsed with sterile saline. The donor heart fragment isplaced into the preformed tunnel with the insertion tool and air orresidual fluid is gently expressed from the tunnel with light pressure.No suturing, adhesive bonding, bandaging, or treatment with antibioticsis required.

Implants are examined at 10-20-fold magnification with a stereoscopicdissecting microscope without anesthesia. Recipients whose grafts arenot visibly beating may be anesthetized and evaluated for the presenceof electrical activity using Grass E-2 platinum subdermal pinmicroelectodes placed either in the pinna or directly into the graft anda tachograph. Implants can be examined 1-4 times a day for 10, 20, 30 ormore days. The ability of a test compound to ameliorate symptoms oftransplant rejection can be compared with a control compound such ascyclosporine, tacrolimus, or orally-administered lefluonomide.

Example 10

Murine Ovalbumin-induced Eosinophilia

Female C57BL/6 mice are obtained from the Jackson Laboratory (BarHarbor, Me.). All animals are given food and water ad libitum. Mice aresensitized with a single i.p. injection of OVA (grade V, Sigma ChemicalCompany, St. Louis, Mo.) adsorbed to alum, (10 μg OVA+9 mg alum in 200μl saline) or vehicle control, (9 mg alum in 200 μl saline) on day 0. Onday 14, the mice are challenged with a 12-minute inhalation of anaerosol consisting of 1.5% OVA (weight/volume) in saline produced by anebulizer (small particle generator, model SPAG-2; ICN Pharmaceuticals,Costa Mesa, Calif.). Groups of eight mice are dosed with oral vehicle(0.5% hydroxypropylmethylcellulose/0.25% TWEEN-80), or a test compoundat 10, 30, or 100 mg/kg in oral vehicle, 200 μl per mouse p.o. Dosing isperformed once per day starting on day 7 or day 13, and extendingthrough day 16.

For determination of pulmonary eosinophilia, three days after the firstOVA aerosol challenge (day 17), the mice are anesthetized with an i.p.injection of anesthetic (Ketamine/Acepromazine/Xylazine), and thetracheae is exposed and cannulated. The lungs and upper airways arelavaged twice with 0.5 ml of cold PBS. A portion (200 μl) of thebronchoalveolar lavage (BAL) fluid is enumerated using a Coulter counterModel ZB1 (Coulter Electronics, Hialeah, Fla.). The remaining BAL fluidis then centrifuged at 300×g for five minutes, and the cells areresuspended in 1 ml of HBSS (Gibco BRL) containing 0.5% fetal calf serum(HyClone) and 10 mM HEPES (Gibco BRL). The cell suspension iscentrifuged in a cytospin (Shandon Southern Instruments, Sewickley, Pa.)and stained by Diff Quick (American Scientific Products, McGraw Park,Ill.) to differentiate BAL leukocytes into neutrophil, eosinophil,monocyte or lymphocyte subsets. The number of eosinophils in the BALfluid is determined by multiplying the percentage of eosinophils by thetotal cell count.

OTHER EMBODIMENTS

From the above disclosure and examples, and from the claims below, theessential features of the invention are readily apparent. The scope ofthe invention also encompasses various modifications and adaptationswithin the knowledge of a person of ordinary skill. Examples include adisclosed compound modified by addition or removal of a protectinggroup, or an ester, pharmaceutical salt, hydrate, acid, or amide of adisclosed compound. Publications cited herein are hereby incorporated byreference in their entirety.

What is claimed is:
 1. A method for treating a proliferative disease,said method comprising administering to a patient in need of suchtreatment a pharmaceutically-effective amount of a compound of formula(I):

R₁ is H, C₁₋₈ a alkyl, C₃₋₈ alkenyl, C₃₋₈ alkynyl, ,C₃₋₄ cycloalkyl,phenyl, (phenyl)C₁₋₄ alkyl, (phenyl)C₃₋₄ alkenyl, (phenyl)C₃₋₄ alkynyl,(C₃₋₈ cycloalkyl)-C₁₋₄ alkyl, (C₃₋₂ cycloalkyl)C₃₋₄ alkenyl, (C₃₋₈cycloalkyl)C₃₋₄ alkynyl, C₃₋₈ heterocyclic radical, (C₃₋₈ heterocyclicradical)C₁₋₄ alkyl, (C₁₋₈ heterocyclic radical)C₃₋₄ alkenyl, (C₃₋₈heterocyclic radical)C₃₋₄ alkynyl, (CH₂)₂₋₄ OR_(C)or (CH₂)₂₋₄NR_(C)R_(D); R₂ is H,C₁₋₄ alkyl, phenyl, C₃₋₆ heterocyclic radical, or(C₃₋₆ cycloalkyl) methyl; each of R₃ and R₄ is independently selectedfrom H, F, NO₂, Br and Cl; R₅ is selected from H and F; R₆ is H, F, Clor CH₃; each of R_(C) and R_(D) is independently selected from H, C₁₋₄alkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, C₃₋₆ cycloalkyl, and phenyl; orNR_(C)R_(D) may be a piperidino, morpholino, or N-(C₁₋₆ alkyl)piperazinoring; wherein each hydrocarbon radical above is optionally substitutedwith between 1 and 3 substituents independently selected from halo,hydroxyl, amino, (amino)sulfonyl, and NO₂; and wherein each heterocyclicradical above is optionally substituted with between 1 and 3substituents independently selected from halo, C₁₋₄ alkyl, C₃₋₆cycloalkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, phenyl, hydroxyl, amino,(amino)sulfonyl, and NO₂, wherein each substituent alkyl, cycloalkyl,alkenyl, alkynyl or phenyl is in turn optionally substituted withbetween 1 and 2 substitutents independently selected from halo, C₁₋₂alkyl, hydroxyl, amino, and NO₂; or a pharmaceutically acceptable saltor C₁₋₈ ester thereof.
 2. A method of claim 1 wherein said proliferativedisease is selected from psoriasis, restenosis, autoimmune disease, andatherosclerosis.
 3. A method for treating a MEK-related cancer, saidmethod comprising administering to a patient in need of such treatment apharmaceutically-effective amount of a compound of Formula (I):

R₁ is H, C₁₋₈ alkyl, C₃₋₈ alkenyl, C₃₋₈ alkynyl, C₃₋₈ cycloalkyl,phenyl, (phenyl)C₁₋₄ alkyl, (phenyl)C₃₋₄ alkenyl, (phenyl)C₃₋₄ alkynyl,(C₃₋₈ cycloalkyl)-C₁₋₄ alkyl, (C₃₋₈ cycloalkyl)C₃₋₄ alkenyl, (C₃₋₈cycloalkyl)C₃₋₄ alkynyl, C₃₋₈ heterocyclic radical, (C₃₋₈ heterocyclicradical)C₁₋₄ alkyl, (C₃₋₈ heterocyclic radical)C₃₋₄ alkenyl, (C₃₋₈heterocyclic radical)C₃₋₄ alkynyl, (CH₂)₂₋₄ OR_(C) or (CH₂)₂₋₄NR_(C)R_(D); R₂ is H, C₁₋₄ alkyl, phenyl, C₃₋₄ cycloalkyl, C₃₋₆heterocyclic radical, or (C₁₋₆ cycloalkyl) methyl; each of R₃ and R₄ isindependently selected from H, F, NO₂, Br and Cl; R₅ is selected form Hand F; R₆ is H, F, Cl, or CH₃; each of R_(C) and R_(D) is independentlyselected from H, C₁₋₄ alkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, C₃₋₆cycloalkyl, and phenyl; or NR_(C)R_(D) may be a piperidino, morpholino,or N-(C₁₋₆ alkyl)piperazino ring; wherein each hydrocarbon radical aboveis optionally substituted with between 1 and 3 substituentsindependently selected from halo, hydroxyl, amino, (amino)sulfonyl, andNO₂; and wherein each heterocyclic radical above is optionallysubstituted with between 1 and 3 substituents independently selectedfrom halo, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl,phenyl, hydroxyl, amino, (amino)sulfonyl, and NO₂, wherein eachsubstituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turnoptionally substituted with between 1 and 2 substitutents independentlyselected from halo, C₁₋₂ alkyl, hydroxyl, amino, and NO₂; or apharmaceutically acceptable salt or C₁₋₈ ester thereof.
 4. A method fortreating brain cancer, breast cancer, lung cancer, ovarian cancer,pancreatic cancer, prostatic cancer, renal cancer, or colorectal cancer,said method comprising administering to a patient in need of suchtreatment a pharmaceutically-effective amount of a compound of Formula(I):

R₁ is H, C₃₋₈ a alkyl, C₃₋₈ alkenyl, C₃₋₈ alkynyl, ,C₃₋₈ cycloalkyl,phenyl, (phenyl)C₁₋₄ alkyl, (phenyl)C₃₋₄ alkenyl, (phenyl)C₃₋₄ alkynyl,(C₃₋₈ cycloalkyl)-C₁₋₄ alkyl, (C₃₋₈ cycloalkyl)C₃₋₄ alkenyl, (C₃₋₈cycloalkyl)C₃₋₄ alkynyl, C₃₋₈ heterocyclic radical, (C₃₋₈ heterocyclicradical)C₁₋₄ alkyl, (C₃₋₈ heterocyclic radical)C₃₋₄ alkenyl, (C₃₋₈heterocyclic radical)C₁₋₄ alkynyl, (CH₂)₂₋₄ NR_(C) or NR_(C)R_(D); R₃ isH,C₁₋₄ alkyl, phenyl, C₃₋₆ heterocyclic radical, or (C₃₋₆ cycloalkyl)methyl; each of R₃ and R₄ is independently selected from H, F, NO₂, Brand Cl; R₅ is selected from H and F; R₆ is H, F, Cl or CH₃; each ofR_(C) and R_(D) is independently selected from H, C₁₋₄ alkyl, C₃₋₄alkenyl, C₃₋₄ alkynyl, C₃₋₆ cycloalkyl, and phenyl; or NR_(C)R_(D) maybe a piperidino, morpholino, or N-(C₁₋₆ alkyl)piperazino ring; whereineach hydrocarbon radical above is optionally substituted with between 1and 3 substituents independently selected from halo, hydroxyl, amino,(amino)sulfonyl, and NO₂; and wherein each heterocyclic radical above isoptionally substituted with between 1 and 3 substituents independentlyselected from halo, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₃₋₄ alkenyl, C₃₋₄alkynyl, phenyl, hydroxyl, amino, (amino)sulfonyl, and NO₂, wherein eachsubstituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turnoptionally substituted with between 1 and 2 substitutents independentlyselected from halo, C₁₋₂ alkyl, hydroxyl, amino, and NO₂; or apharmaceutically acceptable salt or C₁₋₈ ester thereof.
 5. A method fortreating, or ameliorating the sequelac of, a stroke, or heart failure,said method comprising administering to a patient in need of suchtreatment a pharmaceutically-effective amount of a compositioncomprising a compound of Formula (I):

R₁ is H, C₁₋₈ alkyl, C₃₋₈ alkenyl, C₃₋₈ alkynyl, C₃₋₈ cycloalkyl,phenyl, (phenyl)C₁₋₄ alkyl, (phenyl)C₃₋₄ alkenyl, (phenyl)C₃₋₄ alkynyl,(C₃₋₈ cycloalkyl)-C₁₋₄ alkyl, (C₃₋₈ cycloalkyl)C₃₋₄ alkenyl, (C₃₋₈cycloalkyl)C₃₋₄ alkynyl, C₃₋₈ heterocyclic radical, (C₃₋₈ heterocyclicradical)C₁₋₄ alkyl, (C₃₋₈ heterocyclic radical)C₃₋₄ alkenyl, (C₃₋₈heterocyclic radical)C₃₋₄ alkynyl, (CH₂)₂₋₄ OR_(C)or (CH₂)₂₋₄NR_(C)R_(D); R₂ is H,C₁₋₄ alkyl, phenyl, C₃₋₆ cycloalkyl, C₃₋₆heterocyclic radical, or (C₃₋₆ cycloalkyl) methyl; each of R₃ and R₄ isindependently selected from H, F, NO₂, Br and Cl; R₅ is selected from Hand F; R₆ is H, F, Cl or CH₃; each of R_(C) and R_(D) is independentlyselected from H, C₁₋₄ alkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, C₃₋₆cycloalkyl, and phenyl; or NR_(C)R_(D) may be a piperidino, morpholino,or N-(C₁₋₆ alkyl)piperazino ring; wherein each hydrocarbon radical aboveis optionally substituted with between 1 and 3 substituentsindependently selected from halo, hydroxyl, amino, (amino)sulfonyl, andNO₂; and wherein each heterocyclic radical above is optionallysubstituted with between 1 and 3 substituents independently selectedfrom halo, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl,phenyl, hydroxyl, amino, (amino)sulfonyl, and NO₂, wherein eachsubstituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turnoptionally substituted with between 1 and 2 substitutents independentlyselected from halo, C₁₋₂ alkyl, hydroxyl, amino, and NO₂; or apharmaceutically acceptable salt or C₁₋₈ ester thereof.
 6. A method fortreating or reducing the symptoms of xenograft rejection, said methodcomprising administering to an organ transplant, limb transplant, skintransplant, cell(s) transplant, or bone marrow transplant patient apharmaceutically-effective amount of a compound of Formula (I):

R₁ is H, C₁₋₈ a alkyl, C₃₋₈ alkenyl, C₃₋₈ alkynyl, C₃₋₈ cycloalkyl,phenyl, (phenyl)C₁₋₄ alkyl, (phenyl)C₃₋₄ alkenyl, (phenyl)C₃₋₄ alkynyl,(C₃₋₈ cycloalkyl)-C₁₋₄ alkyl, (C₃₋₈ cycloalkyl)C₃₋₄ alkenyl, (C₃₋₈cycloalkyl)C₃₋₄ alkynyl, C₃₋₈ heterocyclic radical, (C₃₋₈ heterocyclicradical)C₁₋₄ alkyl, (C₃₋₈ heterocyclic radical)C₃₋₄ alkenyl, (C₃₋₈heterocyclic radical)C₃₋₄ alkynyl, (CH₂)₂₋₄ OR_(C) or (CH₂)₂₋₄NR_(C)R_(D); R₂ is H, C₁₋₄ alkyl, phenyl, C₃₋₆ C₃₋₆ heterocyclicradical, or (C₃₋₆ cycloalkyl) methyl; each of R₃ and R₄ is independentlyselected from H, F, NO₂, Br and Cl; R₅ is selected from H and F; R₆ isH, F, Cl or CH₃; each of R_(C) and R_(D) is independently selected fromH, C₁₋₄ alkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, C₃₋₆ cycloalkyl, and phenyl;or NR_(C)R_(D) may be a piperidino, morpholino, or N-(C₁₋₆alkyl)piperazino ring; wherein each hydrocarbon radical above isoptionally substituted with between 1 and 3 substituents independentlyselected from halo, hydroxyl, amino, (amino)sulfonyl, and NO₂; andwherein each heterocyclic radical above is optionally substituted withbetween 1 and 3 substituents independently selected from halo, C₁₋₄alkyl, C₃₋₆ cycloalkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, phenyl, hydroxyl,amino, (amino)sulfonyl, and NO₂, wherein each substituent alkyl,cycloalkyl, alkenyl, alkynyl or phenyl is in turn optionally substitutedwith between 1 and 2 substitutents independently selected from halo,C₁₋₂ alkyl, hydroxyl, amino, and NO₂; or a pharmaceutically acceptablesalt or C₁₋₈ ester thereof.
 7. A method for treating osteoarthritis,rheumatoid arthritis, cystic fibrosis, hepatomegaly, cardiomegaly,Alzheimer's disease, a complication of diabetes, septic shock, and viralinfection, said method comprising administering to a patient in need ofsuch treatment a pharmaceutically-effective amount of a compound ofFormula (I):

R₁ is H, C₁₋₈ alkyl, C₃₋₈ alkenyl, C₃₋₈ alkynyl, C₃₋₈ cycloalkyl,phenyl, (phenyl)C₁₋₄ alkyl, (phenyl)C₃₋₄ alkenyl, (phenyl)C₃₋₄ alkynyl,(C₃₋₈ cycloalkyl)-C₁₋₄ alkyl, (C₃₋₂ cycloalkyl)C₃₋₄ alkenyl, (C₃₋₈cycloalkyl)C₃₋₄ alkynyl, C₃₋₈ heterocyclic radical, (C₃₋₈ heterocyclicradical)C₁₋₄ alkyl, (C₃₋₈ heterocyclic radical)C₃₋₄ alkenyl, (C₃₋₈heterocyclic radical)C₃₋₄ alkynyl, (CH₂)₂₋₄ OR_(C) or (CH₂)₂₋₄NR_(C)R_(D); R₂ is H, C₁₋₄ alkyl, phenyl, C₃₋₆ cycloalkyl, C₃₋₆heterocyclic radical, or (C₃₋₆ cycloalkyl) methyl; each of R₃ and R₄ isindependently selected from H, F, NO₂, Br and Cl; R₅ is selected from Hand F; R₆ is H, F, Cl or CH₃; each of R_(C) and R_(D) is independentlyselected from H, C₁₋₄ alkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, C₃₋₆cycloalkyl, and phenyl; or NR_(C)R_(D) may be a piperidino, morpholino,or N-(C₁₋₆ alkyl)piperazino ring; wherein each hydrocarbon radical aboveis optionally substituted with between 1 and 3 substituentsindependently selected from halo, hydroxyl, amino, (amino)sulfonyl, andNO₂; and wherein each heterocyclic radical above is optionallysubstituted with between 1 and 3 substituents independently selectedfrom halo, C₁₋₄ alkyl, C₃₋₆ cycloalkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl,phenyl, hydroxyl, amino, (amino)sulfonyl, and NO₂, wherein eachsubstituent alkyl, cycloalkyl, alkenyl, alkynyl or phenyl is in turnoptionally substituted with between 1 and 2 substitutents independentlyselected from halo, C₁₋₂ alkyl, hydroxyl, amino, and NO₃; or apharmaceutically acceptable salt or C₁₋₈ ester thereof.
 8. A method ofclaim 7, wherein said viral infection is an infection of HIV.
 9. Amethod for treating cancer, said method comprising (a) administering toa patient in need of such treatment, a pharmaceutically-effective amountof a compound of Formula (I):

R₁ is H, C₁₋₈ alkyl, C₃₋₈ alkenyl, C₃₋₈ alkynyl, C₃₋₄ cycloalkyl,phenyl, (phenyl)C₁₋₄ alkyl, (phenyl)C₃₋₄ alkenyl, (phenyl)C₃₋₄ alkynyl,(C₃₋₈ cycloalkyl)-C₁₋₄ alkyl, (C₃₋₈ cycloalkyl)C₃₋₄ alkenyl, (C₃₋₈cycloalkyl)C₃₋₄ alkynyl, C₃₋₈ heterocyclic radical, (C₃₋₈ heterocyclicradical)C₁₋₄ alkyl, (C₃₋₈ heterocyclic radical)C₃₋₄ alkenyl, (C₃₋₈heterocyclic radical)C₃₋₄ alkynyl, (CH₂)₂₋₄ OR_(C) or (CH₂)₂₋₄NR_(C)R_(D); R₂ is H,C₁₋₄ alkyl, phenyl, C₃₋₆ heterocyclic radical, or(C₃₋₆ cycloalkyl) methyl; each of R₃ and R₄ is independently selectedfrom H, F, NO₂, Br and Cl; R₅ is selected from H and F; R₆ is H, F, Clor CH₃; each of R_(C) and R_(D) is independently selected from H, C₁₋₄alkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, C₃₋₆ cycloalkyl, and phenyl; orNR_(C)R_(D) may be a piperidino, morpholino, or N-(C₁₋₆ alkyl)piperazinoring; wherein each hydrocarbon radical above is optionally substitutedwith between 1 and 3 substituents independently selected from halo,hydroxyl, amino, (amino)sulfonyl, and NO₂; and wherein each heterocyclicradical above is optionally substituted with between 1 and 3substituents independently selected from halo, C₁₋₄ alkyl, C₃₋₆cycloalkyl, C₃₋₄ alkenyl, C₃₋₄ alkynyl, phenyl, hydroxyl, amino,(amino)sulfonyl, and NO₂, wherein each substituent alkyl, cycloalkyl,alkenyl, alkynyl or phenyl is in turn optionally substituted withbetween 1 and 2 substitutents independently selected from halo, C₁₋₂alkyl, hydroxyl, amino, and NO₂; or a pharmaceutically acceptable saltor C₁₋₈ ester thereof; and administering a therapy selected fromradiation therapy and chemotherapy.
 10. A method of claim 9, whereinsaid chemotherapy comprises a mitotic inhibitor.
 11. A method of claim10, wherein said mitotic inhibitor is selected from paclitaxel,docetaxel, vincristine, vinblastine, vinorelbine, and vinflunine.